Proteomics

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Rapid and in-depth proteomic profiling of small extracellular vesicles for ultralow samples


ABSTRACT: The form and function of extracellular vesicles (EV) is defined by their proteome. This knowledge is essential to describe and understand EVs, encompassing their marker proteins, capacity as a signalling platform, and utility as diagnostic tools and therapeutic targets. However, EV are low-abundant entities in the secreteome and biological samples, with disease-specific EV proteins often present in low abundance challenging their detection and quantification due to the inherent challenges in dynamic range using mass spectrometry-based strategies. Combined with lack of protein amplification mechanisms, their proteomic studies require upscaling cell cultures or larger volumes of biofluids. Here, we outline high-sensitivity sample preparation and finely tuned LC gradients for DIA to obtain precise and comprehensive proteome of EVs from ultra-low sample amounts (sub-nanogram).

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: David Greening  

PROVIDER: MSV000092668 | MassIVE | Wed Aug 16 06:57:00 BST 2023

REPOSITORIES: MassIVE

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Rapid and in-depth proteomic profiling of small extracellular vesicles for ultralow samples.

Cross Jonathon J   Rai Alin A   Fang Haoyun H   Claridge Bethany B   Greening David W DW  

Proteomics 20231003 11


The integration of robust single-pot, solid-phase-enhanced sample preparation with powerful liquid chromatography-tandem mass spectrometry (LC-MS/MS) is routinely used to define the extracellular vesicle (EV) proteome landscape and underlying biology. However, EV proteome studies are often limited by sample availability, requiring upscaling cell cultures or larger volumes of biofluids to generate sufficient materials. Here, we have refined data independent acquisition (DIA)-based MS analysis of  ...[more]

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